不同繁殖季节绵羊卵巢组织中piRNAs序列特征及表达差异分析

doi:10.16431/j.cnki.1671-7236.2020.07.013

中国畜牧兽医 ›› 2020, Vol. 47 ›› Issue (7): 2081-2092.doi: 10.16431/j.cnki.1671-7236.2020.07.013

不同繁殖季节绵羊卵巢组织中piRNAs序列特征及表达差异分析

张仁森¹, 贺建宁², 魏彩虹¹, 梁小军³, 马青³, 狄冉¹, 储明星¹

1. 中国农业科学院北京畜牧兽医研究所, 农业农村部动物遗传育种与繁殖重点实验室, 北京 100193;
2. 青岛农业大学动物科技学院, 青岛 266109;
3. 宁夏农林科学院动物科学研究所, 银川 750002

收稿日期:2019-12-18 出版日期:2020-07-20 发布日期:2020-07-18
通讯作者: 狄冉(1979-),女,山东泰安人,博士,研究方向:动物遗传育种与繁殖,E-mail:dirangirl@163.com;储明星(1968-),男,安徽贵池人,博士,研究方向:动物遗传育种与繁殖,E-mail:mxchu@263.net E-mail:dirangirl@163.com;mxchu@263.net
作者简介:张仁森(1992-),男,湖南湘西人,硕士,研究方向:动物遗传育种与繁殖,E-mail:1014704865@qq.com
基金资助:
国家自然科学基金项目（31572371）；国家肉羊产业技术体系专项（CARS-38）；中国农业科学院科技创新工程（ASTIP-IAS13）

Characterization and Differentially Expression Analysis of Ovine Ovary piRNAs During Different Reproductive Seasons

ZHANG Rensen¹, HE Jianning², WEI Caihong¹, LIANG Xiaojun³, MA Qing³, DI Ran¹, CHU Mingxing¹

1. Key Laboratory of Animal Genetics, Breeding and Reproduction of Ministry of Agriculture and Rural Affairs, Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100193, China;
2. College of Animal Science and Technology, Qingdao Agricultural University, Qingdao 266109, China;
3. Institute of Animal Science, Ningxia Academy of Agriculture and Forestry Sciences, Yinchuan 750002, China

Received:2019-12-18 Online:2020-07-20 Published:2020-07-18

摘要/Abstract

摘要： 为了分析绵羊卵巢组织中piRNAs的序列特征及不同繁殖季节绵羊卵巢组织中piRNAs的表达差异，本试验采用Solexa测序技术对休情季节（休情期）和发情季节（卵泡期和黄体期）滩羊卵巢组织进行高通量测序，采用生物信息学方法筛选绵羊卵巢中的piRNAs，对piRNAs的长度分布、首位碱基偏好、基因组来源、染色体分布、链特异性、ping-pong循环特征、piRNAs比对重复序列类型进行分析，并对发情季节与休情季节绵羊卵巢之间差异表达的piRNAs进行靶基因预测及其靶基因的GO功能和KEGG Pathway富集分析。结果显示，绵羊卵巢中不同长度的piRNAs首位碱基偏好性不一致；3个时期（休情期、黄体期和卵泡期）的piRNAs均主要比对到基因内含子、编码区及lncRNA区域，而比对到重复序列的piRNAs相对较少；绵羊卵巢piRNAs在染色体上呈不均匀分布；绵羊piRNAs簇存在很强的链特异性，但首位碱基偏好性不明显；3个时期的piRNAs均无明显的ping-pong循环特征。KEGG富集分析发现，差异piRNA靶基因主要富集在RNA运输通路（RNA transport pathway）、嘌呤代谢通路（purine metabolism pathway）、黏着斑通路（focal adhesion）、PI3K-Akt信号通路（PI3K-Akt signaling pathway）、Hippo信号通路（Hippo signaling pathway）等与绵羊繁殖相关的通路。本研究揭示了不同繁殖季节绵羊卵巢组织中的piRNAs序列特征，暗示绵羊卵巢组织中的piRNAs可能通过BAD等基因和黏着斑通路等调控绵羊季节性繁殖，为深入解析绵羊季节性发情分子机制提供一定参考。

关键词: piRNAs; 小RNA测序; 绵羊; 季节性繁殖; 卵巢

Abstract: In order to analyze the sequence features of piRNAs and the expression difference of piRNAs between anestrus and estrus season in ovine ovary,Solexa high-throughput sequencing technology was used in ovary tissue of Tan sheep during the anestrous and estrus season (include follicular and luteal phases).Then piRNAs in ovine ovary were screened,and the length distribution,the first base bias,genomic distribution,chromosome distribution,strand specificity and ping-pong signature of piRNAs and the types of repetitive sequence which piRNAs mapped to were analyzed by bioinformatics methods.The differentially expressed piRNAs (DE piRNAs) in ovary of Tan sheep between anestrus and estrus season were screened and the target gene of DE piRNAs were subjected to GO and KEGG analysis.The results showed that the first base of different size piRNAs in ovine ovary were various.All of the piRNAs from three stages were mainly mapped to intron,coding sequence and lncRNA while little mapped to repeat sequences.The distribution of piRNAs in ovary were uneven among the chromosomes.piRNAs in ovine ovary had strong strand specificity.piRNAs from three stages had no obvious ping-pong signature.The KEGG enrichment analysis showed that the target genes of DE piRNAs were mainly enriched in those pathways that were correlated with ovine reproduction,such as RNA transport pathway,purine metabolism pathway,focal adhesion,PI3K-Akt signaling pathway and Hippo signaling pathway and so on.In conclusion,the characters of piRNAs from ovine ovary during different reproductive stages were revealed,the results suggested that piRNAs in sheep ovarian tissue might participate in regulation of seasonal reproduction through target genes such as BAD and pathways such as focal adhesion,which provided some reference for further analysis of the molecular mechanism of sheep seasonal estrus.

Key words: piRNAs; small RNA sequencing; sheep; seasonal reproduction; ovary

中图分类号:

S814.8

张仁森, 贺建宁, 魏彩虹, 梁小军, 马青, 狄冉, 储明星. 不同繁殖季节绵羊卵巢组织中piRNAs序列特征及表达差异分析[J]. 中国畜牧兽医, 2020, 47(7): 2081-2092.

ZHANG Rensen, HE Jianning, WEI Caihong, LIANG Xiaojun, MA Qing, DI Ran, CHU Mingxing. Characterization and Differentially Expression Analysis of Ovine Ovary piRNAs During Different Reproductive Seasons[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(7): 2081-2092.

参考文献

[1] 刘秋月,胡文萍,贺小云,等.绵羊多羔主效基因FecB高通量检测方法的建立及应用[J].畜牧兽医学报,2017,48(1):39-51. LIU Q Y,HU W P,HE X Y,et al.Establishment of high-throughput molecular detection methods for ovine high fecundity major gene FecB and their application[J].Chinese Journal of Animal and Veterinary Sciences,2017,48(1):39-51.(in Chinese)
[2] GIANTSIS I A,LALIOTIS G P,STOUPA O,et al.Polymorphism of the melatonin receptor 1A (MNTR1A) gene and association with seasonality of reproductive activity in a local Greek sheep breed[J].Journal of Biological Research-Thessaloniki,2016,23(1):2-4.
[3] HE J N,HUANG D W,DI R,et al.Polymorphism of exon 2 of DIO2 gene and its association with seasonal reproduction in sheep[J].Turkish Journal of Veterinary & Animal Sciences,2016,40(2):142-149.
[4] 金慧慧,刘秋月,狄冉,等.Kiss1基因调控哺乳动物繁殖性能的研究进展[J].家畜生态学报,2015,36(8):1-6. JIN H H,LIU Q Y,DI R,et al.Research progress on reproduction regulation of Kiss1 gene in mammals[J].Journal of Domestic Animal Ecology,2015,36(8):1-6.(in Chinese)
[5] 卢璐璐,孙晓笛,储明星,等.绵羊AA-NAT基因mRNA表达与常年发情相关性研究[J].畜牧兽医学报,2015,46(4):542-548. LU L L,SUN X D,CHU M X,et al.Association between AA-NAT mRNA expression and year-round estrus in sheep[J].Chinese Journal of Animal and Veterinary Sciences,2015,46(4):542-548.(in Chinese)
[6] 吕晓曼,苏蕊,张文广,等.绵羊繁殖性状相关基因的研究进展[J].中国畜牧兽医,2012,39(10):185-190. LYU X M,SU R,ZHANG W G,et al.Review on genes involved in reproduction of sheep[J].China Animal Husbandry & Veterinary Medicine,2012,39(10):185-190.(in Chinese)
[7] LI B,HE X,ZHAO Y,et al.Identification of piRNAs and piRNA clusters in the testes of the Mongolian horse[J].Scientific Reports,2019,9(1):5022.
[8] STURM Á,PERCZEL A,IVICS Z,et al.The Piwi-piRNA pathway:Road to immortality[J].Aging Cell,2017,16(5):906-911.
[9] ARAVIN A A,GAIDATZIS D,PFEFFER S,et al.A novel class of small RNAs bind to MILI protein in mouse testes[J].Nature,2006,442(7099):203-207.
[10] ARAVIN A A,NAUMOVA N M,TULIN A V,et al.Double-stranded RNA-mediated silencing of genomic tandem repeats and transposable elements in the D.melanogaster germline[J].Current Biology,2001,11(13):1017-1027.
[11] HAN B W,ZAMORE P D.piRNAs[J].Current Biology,2014,24(16):R730-R733.
[12] TÓTH K F,PEZIC D,STUWE E,et al.The piRNA pathway guards the germline genome against transposable elements[J].Advances in Experimental Medicine and Biology,2016,886:51-77.
[13] MADISON-VILLAR M J,SUN C,LAU N C,et al.Small RNAs from a big genome:The piRNA pathway and transposable elements in the salamander species Desmognathus fuscus[J].Journal of Molecular Evolution,2016,83(3-4):126-136.
[14] OZATA D M,GAINETDINOV I,ZOCH A,et al.Piwi-interacting RNAs:Small RNAs with big functions[J].Nature Reviews Genetics,2019,20(2):89-108.
[15] LIN H,YIN H.A novel epigenetic mechanism in drosophila somatic cells mediated by Piwi and piRNAs[J].Cold Spring Harbor Symposia on Quantitative Biology,2008,73(73):273-281.
[16] PENG J C,LIN H.Beyond transposons:The epigenetic and somatic functions of the Piwi-piRNA mechanism[J].Current Opinion in Cell Biology,2013,25(2):190-194.
[17] SARKAR A,VOLFF J N,VAURY C.piRNAs and their diverse roles:A transposable element-driven tactic for gene regulation?[J].FASEB Journal,2017,31(2):436-446.
[18] IWASAKI Y W,SIOMI M C,SIOMI H.PIWI-Interacting RNA:Its biogenesis and functions[J].Annual Review of Biochemistry,2015,84(1):405-433.
[19] ROUGET C,PAPIN C,BOUREUX A,et al.Maternal mRNA deadenylation and decay by the piRNA pathway in the early drosophila embryo[J].Nature,2010,467(7319):1128-1132.
[20] BARCKMANN B,PIERSON S,DUFOURT J,et al.Auberginei CLIP reveals piRNA-dependent decay of mRNAs involved in germ cell development in the early embryo[J].Cell Reports,2015,12(7):1205-1216.
[21] KIUCHI T,KOGA H,KAWAMOTO M,et al.A single female-specific piRNA is the primary determiner of sex in the silkworm[J].Nature,2014,509(7502):633-666.
[22] DI R,HE J,SONG S,et al.Characterization and comparative profiling of ovarian microRNAs during ovine anestrus and the breeding season[J].BMC Genomics,2014,15(1):899.
[23] HE X,LI B,FU S,et al.Identification of piRNAs in the testes of Sunite and Small-tailed Han sheep[J].Animal Biotechnology,2019,18:1-8.
[24] 殷实,秦文昌,王斌,等.三个阶段牦牛睾丸发育过程中piRNA的鉴定及分析研究[J].畜牧兽医学报,2019,50(11):2235-2243. YIN S,QIN W C,WANG B,et al.Identification and analysis of relevant piRNA during testicular development in yak of three stages[J].Chinese Journal of Animal and Veterinary Sciences,2019,50(11):2235-2243.(in Chinese)
[25] YAN Z,HU H Y,JIANG X,et al.Widespread expression of piRNA-like molecules in somatic tissues[J].Nucleic Acids Research,2011,39(15):6596-6607.
[26] YANG F,XI R.Silencing transposable elements in the Drosophila germline[J].Cellular and Molecular Life Sciences,2017,74(3):435-448.
[27] ROOVERS E F,ROSENKRANZ D,MAHDIPOUR M,et al.Piwi proteins and piRNAs in mammalian oocytes and early embryos[J].Cell Reports,2015,10(12):2069-2082.
[28] HUANG X,FEJES TÓTH K,ARAVIN A A.piRNA biogenesis in Drosophila melanogaster[J].Trends in Genetics,2017,33(11):882-894.
[29] 徐璐.鸡精子发生过程中关键piRNAs及Piwil1基因功能初步研究[D].扬州:扬州大学,2017. XU L.Preliminary study on the biological function of piRNAs and Piwil1 gene during spermatogenesis in poultry[D].Yangzhou:Yangzhou University,2017.(in Chinese)
[30] HAN B W,WANG W,LI C,et al.Noncoding RNA.piRNA-guided transposon cleavage initiates Zucchini-dependent,phased piRNA production[J].Science,2015,348(6236):817-821.
[31] MARIE P P,RONSSERAY S,BOIVIN A.From embryo to adult:piRNA-mediated silencing throughout germline development in Drosophila[J].G3-Genes Genomes Genetics,2017,7(2):505-516.
[32] BAGIJN M P,GOLDSTEIN L D,SAPETSCHNIG A,et al.Function,targets,and evolution of Caenorhabditis elegans piRNAs[J].Science,2012,337(6094):574-578.
[33] BEYRET E,LIU N,LIN H.piRNA biogenesis during adult spermatogenesis in mice is independent of the ping-pong mechanism[J].Cell Research,2012,22(10):1429-1439.
[34] LEE H C,GU W,SHIRAYAMA M,et al.C.elegans piRNAs mediate the genome-wide surveillance of germline transcripts[J].Cell,2012,150(1):78-87.
[35] WATANABE T,TOMIZAWA S,MITSUYA K,et al.Role for piRNAs and noncoding RNA in de novo DNA methylation of the imprinted mouse Rasgrf1 locus[J].Science,2011,332(6031):848-852.
[36] KELLEY D,RINN J.Transposable elements reveal a stem cell-specific class of long noncoding RNAs[J].Genome Biology,2012,13(11):R107.
[37] KIMURA M,ISHIDA K,KASHIWABARA S,et al.Characterization of two cytoplasmic poly(A)-binding proteins,PABPC1 and PABPC2 in mouse spermatogenic cells[J].Biology of Reproduction,2009,80(3):545-554.
[38] XU K,YANG L,ZHAO D,et al.AKAP3 synthesis is mediated by RNA binding proteins and PKA signaling during mouse spermiogenesis[J].Biology of Reproduction,2014,90(6):119.
[39] WU Y,XU K,QI H.Domain-functional analyses of PIWIL1 and PABPC1 indicate their synergistic roles in protein translation via 3'-UTRs of meiotic mRNAs[J].Biology of Reproduction,2018,99(4):773-788.
[40] 李华振,狄冉,刘秋月,等.类视黄醇X受体基因调控动物季节性发情和发情周期的研究进展[J].畜牧兽医学报,2019,50(8):1525-1535. LI H Z,DI R,LIU Q Y,et al.Research progress on the regulative role of retinoid X receptors gene in animal seasonal estrus and estrous cycle[J].Chinese Journal of Animal and Veterinary Sciences,2019,50(8):1525-1535.(in Chinese)
[41] 夏青,张效生,刘秋月,等.EYA3和TSHβ在季节性发情和常年发情绵羊中的表达模式[J].畜牧兽医学报,2018,49(2):263-269. XIA Q,ZHANG X S,LIU Q Y,et al.Expression patterns of EYA3 and TSHβ in seasonal estrous and year-round estrous sheep[J].Chinese Journal of Animal and Veterinary Sciences,2018,49(2):263-269.(in Chinese)
[42] IKEGAMI K,YOSHIMURA T.Comparative analysis reveals the underlying mechanism of vertebrate seasonal reproduction[J].General and Comparative Endocrinology,2016,227:64-68.
[43] CAO X H,WANG X Y,LU L L,et al.Expression and functional analysis of the BCL2-associated agonist of cell death (BAD) gene in the sheep ovary during the reproductive cycle[J].Frontiers in Endocrinology,2018,9:512.
[44] JEOUNG M,BRIDGES P J.Cyclic regulation of apoptotic gene expression in the mouse oviduct[J].Reproduction Fertility and Development,2011,23(5):638-644.
[45] 储明星,冯涛,王西筠,等.哺乳动物繁殖季节性相关基因的研究进展[J].中国畜牧兽医,2010,37(6):107-110. CHU M X,FENG T,WANG X Y,et al.Advances on related genes with reproductive seasonality in mammals[J].China Animal Husbandry & Veterinary Medicine,2010,37(6):107-110.(in Chinese)
[46] HE X Y,ZHANG Z Z,LIU Q Y,et al.Polymorphisms of the melatonin receptor 1A gene that affects the reproductive seasonality and litter size in Small-tail Han sheep[J].Reproduction in Domestic Animals,2019,54(10):1400-1410.
[47] JIN X,VON GALL C,PIESCHL R L,et al.Targeted disruption of the mouse Mel(1b) melatonin receptor[J].Molecular and Cellular Biology,2003,23(3):1054-1060.
[48] 肖迪,刘俊,郑桃云.交泰丸对睡眠剥夺小鼠视交叉上核生物钟基因Clock及Bmal1表达的影响[J].湖北中医药大学学报,2019,21(4):25-29. XIAO D,LIU J,ZHENG T Y.Effects of Jiaotai pills on the expression of Clock and Bmal1 in the suprachiasmatic nucleus of sleep deprived mice[J].Journal of Hubei University of Traditional Chinese Medicine,2019,21(4):25-29.(in Chinese)

不同繁殖季节绵羊卵巢组织中piRNAs序列特征及表达差异分析

Characterization and Differentially Expression Analysis of Ovine Ovary piRNAs During Different Reproductive Seasons

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价

[1]	王赛桥, 赵路, 翟振翰, 张丙雷, 贾万航, 王玉琴. 绵羊miR-449a/b前体序列组织表达及生物信息学分析[J]. 中国畜牧兽医, 2023, 50(6): 2175-2184.
[2]	刘锦钰, 张杨, 马旭梅, 周千, 周扬, 张春香, 石磊, 任有蛇. 谷胱甘肽对绵羊精液低温保存效果的影响[J]. 中国畜牧兽医, 2023, 50(6): 2388-2394.
[3]	崔双, 乌日力嘎, 阿拉达尔, 晓敏, 王剑秋, 弓剑. 放牧条件下补充酵母硒对绵羊胃肠道微生物区系的影响[J]. 中国畜牧兽医, 2023, 50(3): 952-965.
[4]	马建青, 杨清芳, 宋占锋, 贾丁丁, 吴东蔚, 莫秀芳, 徐增年, 杨克, 吴占勇, 籍颖, 周荣艳. 绵羊骨代谢相关基因VDR密码子偏好性分析[J]. 中国畜牧兽医, 2022, 49(8): 2888-2898.
[5]	张雪萍, 石斌刚, 金夏阳, 王向彦, 兰丽娟, 时钰, 祁有鹏, 赵世杰, 李少斌, 胡江. 藏绵羊KLF7基因表达特征分析及其过表达对前脂肪细胞增殖分化的影响[J]. 中国畜牧兽医, 2022, 49(8): 2920-2930.
[6]	李卫东, 蒋玉婷, 段宏伟, 何海军, 杨帅, 丁自强, 吴建新, 张恩秋, 胡俊杰. 褪黑素受体MT1和MT2在绵羊胃中的分布及表达[J]. 中国畜牧兽医, 2022, 49(8): 3256-3262.
[7]	严啊妮, 沈奔, 王洪荣, 冯春燕, 魏宗友. 过瘤胃葡萄糖和缓释尿素对热应激绵羊生产性能及瘤胃发酵功能的影响[J]. 中国畜牧兽医, 2022, 49(6): 2105-2115.
[8]	胡文, 霍孔林, 宋星星, 张鑫, 张多妮, 罗荣荣, 徐文镐, 李珣. 促性腺激素抑制激素对SD大鼠性腺生殖功能与糖代谢的影响[J]. 中国畜牧兽医, 2022, 49(6): 2239-2249.
[9]	张丽萌, 刘爱菊, 李闰婷, 李玉华, 李林, 聂晓宁, 王林青, 陈龙欣. 绵羊CREBRF基因克隆、生物信息学及组织表达分析[J]. 中国畜牧兽医, 2022, 49(5): 1599-1609.
[10]	强浩, 梁鹏, 孟科, 荣轩, 冯登侦. 绵羊IGF1R基因多态性及其与生长性状的关联分析[J]. 中国畜牧兽医, 2022, 49(5): 1765-1785.
[11]	苏晓慧, 何海迎, 吐尔逊阿依·牙力坤, 王琼, 马海玉, 刘玲玲, 刘武军. 野生盘羊×巴什拜羊杂交二代尾部脂肪差异表达lncRNA的鉴定与分析[J]. 中国畜牧兽医, 2022, 49(5): 1796-1805.
[12]	田广原, 王宇琛, 周雅坪, 郭婷, 赵红梅, 赵逢淼, 孙雅婕, 卞宇晨, 于佳梁, 周雨霞. 绵羊肺炎支原体P60_58aa-928aa蛋白的原核表达及间接ELISA方法的建立[J]. 中国畜牧兽医, 2022, 49(5): 1960-1969.
[13]	翟哲, 陈思, 吴艳茹, 王雪梅, 李崇瑞, 刘志勇, 陈巧玲, 王凤阳, 杜丽, 李昌, 金宁一. 绵羊CCL19基因启动子活性分析及其转录调控元件筛选[J]. 中国畜牧兽医, 2022, 49(4): 1213-1222.
[14]	李静云, 王新乐, 张云飞, 白俊艳, 李淦, 雷莹, 董智豪, 陈宇, 樊红灯, 王龙威, 陈梦柯, 王兴科, 司马磊阳, 张世文. 绵羊MyoG基因内含子Ⅱ多态性及其与生长性状的关联分析[J]. 中国畜牧兽医, 2022, 49(4): 1364-1373.
[15]	石雨竹, 罗儒唐, 陈超, 吴旭, 李昂. 番鸭GDF9基因克隆、生物信息学及组织表达分析[J]. 中国畜牧兽医, 2022, 49(3): 817-829.